Genetic Characterization Of Full-length Human Immunodeficiency Virus Type 1 Genomes From Three Infected Paid Blood Donors In Central China And Construction Of Infectious Molecular Clones

We have previously demonstrated that there are two major HIV-1 subtypes in China, subtype B' in the former paid blood donors (PBD) in Henan, and CRF07 and 08 in the IDU in Yunnan, Guangxi and Xinjiang. We have now extended our previous studies by performing BootScan analysis on full-length genome from Henan strains from PBD, revealed the first complete genome background of the most dominant circulating HIV-1 strain in Henan and Hubei, found that HIV in Henan has not yet recombined with virus from those circulating in other area in China and other subtypes around the world. Later, we systematically characterized this three full-length HIV-1 genomes and contructed the first infectious molecular clones of HIV-1 B' from PBD in Henan in 2002. We have found that the three novel clones cluster tightly among themselves and are genetically closest to the HIV-1 B' strain previously identified in Yunnan and Henan without apparent recombination with any other circulating subtypes in China, nicely comfirmed our previous conlusion, and suggesting the possibility of a common origin of a single founder virus before they spreading out to other places in the current affecting region, since the three viral genomes although all from Henan but came from three geographic distinct locations. Two of the three clones have complete open reading frames for all known viral proteins and functional domains and no drug genetic mutations were found in the protease and reverse transcriptase genes, consistent with the time frame when samples were taken before nation-wide free antiretroviral treatment had begun. Many of the epitopes recognized by the potent neutralizing antibodies are also invariant. Using the 3 full-length genomes from PBD in Henan, we constructed 6 infectious HIV-1 subtype B' molecular clones. Among the 6 clones, two clones are from patient 02HNscl 1, the other 4 are chimeric molecules, because they areall consist of genes from different patients. All 6 clones are R5-tropism virus, can effectively infect TZM-bl cells and replicate in CEMxl74-5.25M7 cells, and 5 were able to replicate in PBMC. To our knowledge, we constructed the first infectious HIV-1 IT molecular clone from China. We believe that information provided in this study will have important implications for future design and development of an effective vaccine and antiretroviral drugs targeted at the HIV-1 strains circulating in central China. The infectious clones will be important tools for identifying correlates of protection, epitopes that may confer resistance to infection or disease, as well as for facilitating the test of potential vaccines or drugs against HIV-1 strains circulating in central China.